Site specificity of psoralen-DNA interstrand cross-linking determined by nuclease Bal31 digestion
- 1 October 1986
- journal article
- research article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 25 (21), 6598-6603
- https://doi.org/10.1021/bi00369a039
Abstract
A novel method for determination of psoralen photo-cross-linking sites in double-stranded DNA is described, which is based on a pronounced inhibition of Bal31 exonuclease activity by psoralen-DNA interstrand cross-links. The results using a 51 base pair fragment of plasmid pUC19 and a 346 base pair fragment of pBR322 show that 5''-TA sequences are preferred cross-linking sites compared to 3''-TA sequences. They also indicate that sequences flanking the 5''-TA site influence the cross-linking efficiency at the site. The DNA photo-cross-linking by 4,5'',8-trimethylpsoralen and 8-methoxypsoralen was analyzed, and these two psoralens showed identical site specificity. The 5''-TA preference is rationalized on the basis of the local DNA structure in terms of the .pi.-.pi. electronic interaction between the thymines and the intercalated psoralens, as well as on the base tilt angles of the DNA.This publication has 15 references indexed in Scilit:
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